What happened on Days 2 & 3 was that there was a serious yeast contamination in the cultures set up! Why this happens: Sometimes, the bacterial strain loses its ability to make the drug 9DS: The genes that make 9DS are turned down. Because 9DS fumagates the culture and keeps fungi from growing, when this happens, yeast can contaminate the media and grow.

Day 2:

2400: Resuspend B. subtilis picked from LB plate (Teknova) in 300 uL distilled water, let sit for ~5 minutes to denature the bacterial biofilm. Plate the solution onto a new LB plate and core 6 cores using the wide end of a 1 mL pipette tip. Into each core, deposit 100 uL of culture from respective tubes. Let grow overnight and observe. Culture 5 was very obviously contaminated with fungal growth due to pungent smell and bright yellow color, and was disposed.

Day 3: Observation of plate showed no positive cultures.

2400: Resuspend B. subtilis picked from LB plate (Teknova) in 300 uL distilled water, let sit for ~5 minutes to denature the bacterial biofilm. Plate the solution onto a new LB plate and core 5 cores using the wide end of a 1 mL pipette tip. Into each core, deposit 100 uL of culture from respective tubes. Let grow overnight and observe.

Day 4:

Observation of plate showed no positive cultures.

Up next: How to ‘reboot’ these bacteria and get them to start making 9DS again.

As a part of winding down phase I of Project Marilyn, I’m in the process of liquidating some of the assets of indysci to pay off some liabilities (namely, the cost of the xenograft experiment). As you can see, budget transparency is an important part of indysci, and so it’s time to sell off the bitcoin that was raised during the initial fundraising. I attempted to do this several months ago when signing the contract with the xenograft CRO (in anticipation of having to spend it then), but ran into some roadblocks, since it appeared coinbase was transitioning its requirements for merchant accounts. Because the CRO wanted payments in installments, I figured it would be OK to wait a bit.

Now, the time is coming close to pay for the xenograft, and I sent in an inquiry to get my nonprofit’s bank account approved. Instead, coinbase opted to suspend indysci’s account, “to comply with FinCEN”. They offered no further explanation.

So I will be looking for another place to liquidate indysci’s bitcoin. I chose to take donations in bitcoin in part because I believe that noninflationary currencies are better for global sustainability and better for charitable organizations than an asset that loses value over time. At the time, Coinbase was the easiest way to set up a donation link. Coinbase also provided a nice API that let me to show in real-time the value of the donations to that date, as a way to integrate it into my (Tilt open) crowdfunding. Unfortunately, I can no longer recommend this service for nonprofits: If you’re a nonprofit, don’t use Coinbase to collect donations.

As phase I of Project Marilyn is winding down (waiting on the results from the xenograft experiment), I moved out of the lab at QB3 in San Francisco. As part of this project is to show how easy it can be to make this drug, I decided to actually do all the steps at home. I’ll also take a careful log of everything I’m doing along the way. Here’s day zero of the process.